Make Your Own Mate & Plate Library System

Library generation using n vivo recombination in S. cerevisiae. Mate & Plate Libraries are created via recombination between your cDNA and the Matchmaker prey vector pGADT7-Rec.

Library generation using in vivo recombination in S. cerevisiae. Mate & Plate Libraries are created via recombination between your cDNA and the Matchmaker prey vector pGADT7-Rec. The homologues are generated by SMART cDNA synthesis. Colonies are pooled, mixed, and aliquoted into multiple vials. Each single 1 ml vial can be used for a two-hybrid screen.

Mate & Plate libraries display broad insert representation

Mate & Plate libraries display broad insert representation. A human bone marrow library was made using the Make Your Own “Mate & Plate” Library System. Inserts from 15 randomly picked colonies were analyzed by yeast colony PCR using the Advantage 2 Polymerase Mix (Cat. No. 639201), and the Matchmaker AD LD-Insert Screening Amplimer Set (Cat. No. 630433). As seen in Lanes 1–15, every colony contained an insert of a different size. Lane M: 1 kb DNA ladder molecular weight marker.

SMART cDNA synthesis generates cDNA ends with homology to pGADT7-Rec

SMART cDNA synthesis generates cDNA ends with homology to pGADT7-Rec.